Pyruvaldehyde-bis-(4-methyl-3-thiosemicarbazones) in a trichomonacidal method



United States Patent 3265 s70 PYauvALnnnYnE-nrs- 4 METHYL s rnrosrnvn- CARBAZONES) IN A TRICHOMONACIDAL METHOD Rhoda M. Michaela, Kalamazoo, MiclL, assignor to The Upjohn Company, Kalamazoo, Mich, a corporation of Michigan No Drawing. Filed Dec. 5, 1960, Ser. No. 73,536 7 Claims. (Cl. 16758) This invention relates to therapeutic compositions containing, as the primary active ingredient, certain pyruvaldehyde-bis(4-methyl-3-thiosemicarbazones) for the treatment of trichornonal infections in humans and in cattle and to a method for the use of such compositions.

The primary active ingredients of these compositions are pyruvaldehyde-bis(4-methy1-3-thiosernicarbazones) of the formula:

Patented August 9, 1966 "ice subcutaneously with T. vaginalis a substantial spread is indicated between the curative dose and the tolerated dose. Groups of 5 adult male albino mice, Mus. musculus, were injected subcutaneously (left side) with 200,000 24-hour cells, contained in 0.2 m1. Chemicals for treatment of the resulting infections were homogenized in 2.5% w./v. aqueous gelatin solution. Starting with the tolerated dose (the maximum amount of drug in mg./kg./day which can be administered for six consecutive days and still allow weight gains comparable to control animals), the compounds on test were titrated in vivo by serial twofold dilutions. The first treatment was given immediately after infection, and treatment was continued once daily for five additional days. 'Efiicacy was evaluated by the oral and intraperitoneal routes of treatment. Controls were sham-dosed with 2.5% w./v. aqueous gelatin solution. Since the pathogenicity of the cultures varied, a percent infective dose titration (ID was made for each individual experiment. Serial threefold and serial four-fold dilutions were used for T. vaginalis and T. foetus, respectively. Animals were necropsied seven days after infection. The presence of trichomonads in the lesions which formed at the site of injection were verified by microscopic examination of an eosin-stained wet smear. The 50 percent curative dose (CD and the ID were calculated according to the method of Reed and Muench, Amer. J. Hyg. 27: 493 (1938).

Pertinent data are summarized in the following table, which includes also a significant comparison with a, pair of anti-trichomonal agents currently receiving attention in the literature:

InVitro, meg/m1. Tolerated Dose In Vivo, CD mgJkgJday Oral, mgJkg.

Primary Active Ingredient T. vogi'nalis T. foetus T. T. foetus LP. Oral voginalis LP. Oral LP. Oral Pyruvaldehyde-bis(4-methyl-3-thiosemlcarbaz0ne) 0. 005 0. 005 50 800 0. 33 0. l6 5 2 Hydroxypyruvaldehyde-bis(4-methyl-3-thiosemicarazone)--- 0. 002 0. 009 50 800 0. 33 0. 1. 37 2. 0 Ethoxypyruvaldehyde-bis(4-methyl-3-thiosemiearbazone) 0. 003 0. 06 50-100 600 0. 38 0. 1. 6 2. 1 Ethoxy-fl-methyl-pyruvaldehyde-bis(4-methyl-3-thiosemiearbazone) 0. 0006 0.01 100 800 0. 55 0. 2-Acetylamin0-5-nitrothiazole (Tritheon) 0.62 0. 62 11. 6 11. 6 (Hydroxy-2-ethy1)-1-methy1-2Pnitr0-5-imidazole (Flagyl). 2. 5 2500 2.12

Vaginitis originating with the parasitic protozoan Trichomonas vaginalis is an infection of the human vagina occurring in many millions of women throughout the world. Although the organism is likewise found not infrequently in the genito-urinary tract of the male, its presence there does not give rise to overt disease. Numerous approaches to the treatment of this troublesome afliiction in the female have been tried, but none are without limitations of safety, efficacy or permanence of cure which circumscribe their usefulness. Major emphasis has heretofore been placed on topical treatment of the infection. However, evidence is accumulating that in the female the parasite localizes in such inaccessible sites as Skenes ducts, Bartholins glands and in the urinary tract and other vicinal glands and ducts, thus making it extremely difficult to eliminate the infection by local treatment alone.

It has now been shown that the active ingredients hereof are outstandinglyeffective against T. vaginalis and Trichomonas foetus. On administration to mice infected It is seen from the data above that the primary active ingredients have demonstrated both striking activity and a satisfactory margin of safety when given orally. These properties make possible the oral treatment of T. vaginalis infections in humans, including male carriers, and of T. foetus infections in animals, such as cattle, and other mammals susceptible to the infection. Because of similarly high potency when applied in a topical formulation, a dual systemic and topical regimen can be advantageously employed where appropriate.

Such oral and topical compositions can include supplementary active ingredients which extend the usefulness of the basic formulations or accelerate the desired clinical response to therapy. Of particular interest in this regard are antimonilial agents, such as nystatin, amphotericin B, trichomycin and the like.

The compositions of the present invention are suitably presented for administration in a'variety of unit dosage domestic animals. Suitable oral dosage forms include tablets, pills, capsules, granules, powders, oral solutions or suspensions and the like. For preparing solid compositions such as tablets, the active ingredient is admixed with conventional tableting ingredients or pharmaceutical carriers such as cornstarch, lactose, dicalcium phosphate, talc, stearic acid, magnesium stearate, gums and functionally similar materials serving as pharmaceutical diluents or carriers. The tablets or pills of these novel compositions can be laminated or otherwise compounded to provide a dosage form aifording the advantages of prolonged, delayed or predetermined successive action of the enclose medication. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope enclosing the former. Two components can be separated by an enteric layer which serves to resist disintegration until the desired site of release is encountered. A variety of materials can be used for such enteric layers or coatings, .these materials including a number of polymeric acids or mixtures of polymeric acids with, for example, shellac, shellac and cetyl alcohol, cellulose acetate phthalate (as described in US. Patent No.

2,196,768), and the like. Advantageously, a coating consisting essentially of gelatin and styrene-maleic acid copolymer, deposited on the drug particles by a process of phase separation,-such as by coacervatiou, can be employed as the encapsulating material capable of affording sustained release of the enclosed medication.

The oral liquid forms in which the novel compositions of this invention can be incorporated include aqueous solutions, flavored syrups, 'elixirs, suspensions, flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil, peanut oil, and the like. Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, acacia, alginates, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, gelatin and the like. A particularly useful suspending vehicle is prepared from a combination of sodium carboxymethylcellulose and polyvinylpyrrolidone as disclosed in US. Patent No. 2,897,170. For veterinary use an aqueous suspension prepared as above and sterilized offers a convenient form for administration by injection.

Topical compositions incorporating the primary active ingredients hereof are those prepared in such conventional pharmaceutical forms as ointments, lotions, pastes, jellies, powders, suppositories and the like for vaginal use. Such topical compositions typically include, in addition to the primary and supplementary active ingredients, such materials as emulsifying agents, solvents, antioxidants, preservatives, butfers, perfumes and bodying materials which confer on the product a desired consistency rendering it adaptable to topical use.

The term unit dosage form as applied to the compositions of this invention refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active material intended to produce the desired therapeutic effect in association with the required pharmaceutical diluent, carrier or vehicle. The specifications for the novel unit dosage forms of this invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active material for therapeutic use, as disclosed in detail herein, these being features of the present invention. Examples of suitable unit dosage forms in accord with this invention are tablets, capsules, pills, powder packets, wafers, cachets, teaspoonfuls, tablespoonfuls, dropperfuls, segregated multiples of any of the foregoing, vaginal tablets, vaginal suppositories, and other forms as hereinbefore described.

In the oral treatment of T. vaginalz's vaginitis infection in the adult female, compositions containing from about 15 to about 1000 mg. of the selected pyruvaldehydebis(4-methyl-3-thiosemicarbazone) are given one to three times daily, from about 75 to about 225 mg. three times daily being preferred. The foregoing dosage schedule is likewise applicable where it is desired to treat the male adult as a carrier of the infection. In the oral treatment of T. foetus infections in cattle, a dose of from about 1 to about 10 gm. daily, preferably about 5 gm., is employed. Medication can be given in a bolus or as a food supplement or pre-mix. Parenteral forms are administered in amounts of about one-half those used orally. In both human and veterinary use, however, the exact dosage must ultimately be determined individually in the light of the subjects age, weight and physical condition and the severity of the infection being treated.

Because the said primary active ingredients are likewise etfective when applied topically at the site of the infection, the present compositions contemplate also those pharmaceutical forms adapted to administration by the vaginal route. In particular, powders, ointments, jellies, fluid aqueous or :oil suspensions, tablets and suppositories offer convenient means of treatment for both human and veterinary use. For human vaginal administration, powder, ointment and jelly compositions containing from about 0.0 1 to about 5% of primary active ingredient can be employed, the preferred range [being from about 0.1 to about 2.5%. In general, single applications of such pharmaceutical forms should represent from about 1 to about 250 mg. of principal active ingredient. Vaginal tablets and suppositories suitably contain from about 1 to about 250mg. of primary active ingredient, from about 10 to about mg. being preferred. Higher concentrations or doses can be employed where indicated by the patients age, weight, condition and severity of the infection. In the veterinary .use of these compositions, commonly employing an ointment or jelly as the most convenient form, the same concentrations of active ingredients are used but the quantity applied is suitably about ten times that used in treatment of humans. Topical treatment with compositions such as those indicated above consists in the usual case of administration of a single dosage unit twice daily, e.g., morning and night.

A particularly effective clinical approach employing the compositions of this invention resides in concomitant oral and vaginal therapy. Thus, the human female is placed on a regimen of oral treatment which is supple mented by administration of the compositions in appropriate form by the vaginal route. Some allowance in establishing the oral dosage schedule may be desirable in view of the collateral amounts being given intrav-aginally, although full dosages by both routes can be maintained in most instances.

The following examples set forth the best mode contemplated by the inventor for carrying out this invention, but they are not to be construed as limiting the scope thereof.

EXAMPLE 1.TABLETS A lot of 10,000 tablets for oral use, each containing The finely powdered materials are thoroughly mixed and granulated with 10% starch paste. The dry granules are mixed with a lubricant mixture of 100 gm. of bolted starch and 20 gm. of calcium stearate and then compressed into tablets. These tablets are suitable for clinical use, preferably on a schedule of one tablet daily.

EXAMPLE 2.HARD-GELATIN CAPSULES A lot of 1 0,000 two-piece hard-gelatin capsules for oral use, each capsule containing 75 mg. of hydroxypyruvaldehyde-bis(4-methyl-3-thiosemicarbazone), is prepared from the following materials:

- Hydroxypyruvaldehyde-bis 4-methyl- 3- thiosemicarbazone) 750 Cornstarch 1600 Light mineral oil 130 Magnesium stearate, powder 160 Talc 160 The powdered ingredients are thoroughly mixed and encapsulated in the usual manner. The resulting capsules are suitable for clincical use in a dosage of one capsule daily.

Substitution of 150 gm. of active ingredient for the 750 gm. above gives capsules each containing 15 mg. thereof which can be administered on a schedule of one capsule three times daily.

EXAMPLE 3.+SOFT-ELASTIC CAPSULES A lot of 10,000 soft-elastic capsules for oral use, each containing 225 mg. of ethoxypyruvaldehyde-bis(4-methyl- 3-thiosemicarbazone), is prepared by dispersing 2250 gm.

of the said ingredient in suflicient corn oil to render the drug capsulatable and thereafter forming soft-elastic capsules in the usual manner. One capsule daily represents a suitable dosage schedule.

Substitution of 10,000 gm, of active ingredients for the 2250 gm. above gives capsules each containing 1000 mg. thereof which can be administered on a schedule of one capsule daily.

EXAMPLE 4.-AQUEOUS SUSPENSION Ten liters of aqueous suspension for oral use containing in each 5 ml. 15 mg. of ethoxy-fl-methylpyruvaldehydebis-(4 methyl-3-thiosemica-rbazone) is prepared by thor- .oughly mixing together the following materials: Ethoxy-fi-methylpyruvaldehyde-bi-s(4-methyl- 3-thiosemicarbazone), gm. 30 Methylparaben, gm. 7.5 Propylparaben, gm. 2.5 Saccharin sodium,,gm. 12.5 Cyclamate sodium (sodium cyclohexylsulfamate), gm. 2.5

Glycerin, ml. 3000 Tragacanth powder, gm. Orange oil flavor, gm. 10 RD. and C. orange dye, gm. 7.5

Deionized water, q.s., 10,000 ml.

The suspension prepared as above is suitable for clinical use and can be administered on a dosage schedule of l teaspoonful (5 ml.) given 3 times daily.

EXAMPLE 5.-CAPSULES Ten thousand hard gelatin capsules, each containing 125 mg. of pyruvaldehyde-bis(4-methyl-3-thiosemicarbazone) and 250,000 units of nystatin, are prepared from the following materials:

Gm. Pyruvaldheyde-bis(4-methyl-3- thiosemicarbazone) 1,250 Nystatin (2000 unit/mg.) 1,250 White mineral oil 65 Magnesium stearate, powdered 65 Talc 65 The materials are thoroughly mixed and capsulatcd in the usual manner. The capsules are suitable for administration on a schedule of one capsule given three times daily.

6 EXAMPLE 6.VAGINAL TABLETS One thousand vaginal tablets, each tablet containing mg. of ethoxypyruvaldehyde-bis (4-methyl-3-thiosemicarbazone), are prepared from the following materials:

The ethoxypyruvaldehyde-bis(4-methyl-3-thiosemicarbazone), lactose and alginic acid of Part I are mixed with a portion of the starch of Part I. With the balance of the starch a 10% starch paste is formed for preparation of the granulation. To the dried granules constituting Part I is added the mixture comprising Part II (lubricant) and the resulting granulation is compressed into tablets. The tablets thus prepared are suitable for intravaginal use, preferably one tablet administered twice daily.'

EXAMPLE 7.VAGINAL SUPPOSITORIES A vaginal suppository is prepared by molding a mixture of the following ingredients into suppositories weighing 1 gm. each: 7

Pyruvaldehyde-bis(4-methyl-3- thiosemicarbazone),micronized 10 0 Polyethylene glycol 6000 600 Lactose 300 Suppositories prepared as above are suitable for intravaginal administration, preferably twice daily.

EXAMPLE 8.VAGINAL JELLY A vaginal jelly containing 10 mg. of hydroxypyruvaldehyde-bis(4-methyl-3-thiosemicarbazone) per gm. (1% by weight) is prepared by thoroughly mixing together the following ingredients:

' Y Gm. Hydroxypyruvaldehyde-bis(4-methyl-3- thiosemicarbazone), micronized 10 Polysorbate 8O 14 Polyethylene glycol 400 50 Sodium carboxymethylcellulose 20 Sodium chloride 8 Sodium benzoate 3 Water, q.s., 1000 ml.

The resulting jelly is applied locally to the affected area, preferably twice daily with a 2-ml. applicator.

For treatment of T. foetus infections in cattle, the foregoing jelly can be applied intravaginally in 20-ml. portions.

EXAMPLE 9.-VETERINARY PRE-MIX Ten thousand grams of a pre-mix is prepared by thoroughly mixing 1000 gm. of pyruvaldehyde-bis(4- methyl-3-thiosemicarbazone) into 9000 gm. of soybean meal. Each gram of the pre-mix thus contains 100 mg. of active ingredient. The pre-mix is added to the standand ration of cows to provide a daily dose of 10 gm. of active ingredient for the treatment of T. foetus infections.

I 7 EXAMPLE 10.VETERINARY BOLUS Ten thousand boluses, each containing 5 gm. of hydroxypyruvaldehyde bis (4-methyl-3-thiosemicarbazone) are prepared from the following ingredients:

Hydroxypyruvaldehyde-bis (4-methyl-3 thiosemicarbazone) 50,000 Lactose 250,000

The above ingredients are blended and granulated with syrup-starch paste, and q.s. mineral oil is added. The granulation is then dried, lubricated with starch, talc and calcium stearate powders, and compressed with a 1 /2" X die. One bolus daily is given to cows for treatment of T. foetus infections! EXAMPLE 1 1 wherein R is a member selected from the group consisting of lower alkyl of 1 through 6 carbonatoms, hydroxymethyl, ethoxymethyl and u-ethoxyethyl, dispersed in a solid pharmaceutical carrier.

2. A method for treating Trichomonas vaginalis and Trichomonas foetus infections in humans and cattle which comprises: orally administering a composition containing, as the primary active ingredient, from about 15 to about 1000 mg. of a compound of the formula wherein R is a member selected from the group consisting of lower alkyl of 1 through 6 carbon atoms, hydroxymethyl, ethoxymethyl and a-ethoxyethy l, dispersed in a solid pharmaceutical carrier.

3. A method for treating Trichomonas vaginalis and T richomonas foetus infections in humans and cattle which comprises: orally administering a composition containing, as the primary active ingredient, from about 75 to 0 about 225 mg. of pyruvaldehyde-bis(4-methyl-3-thiosemi- 'carbazone), dispersed in a solid pharmaceutical carrier.

4. A method for treating Trichomonas vaginalis and Trichomonas foetus infections in humans which com prises: topically administering a composition containing, as the primary active ingredient, from about 0.01 to about 5 of pyruvaldehyde-bis(4-methyl-3-thiosemicarbazone dispersed in a topical pharmaceutical carrier.

5. A method for treating T richomonas vaginalis infections in humans which comprises: intravaginally administering a composition containing, as the primary active ingredient, from about 1 to about 250 mg. of pyruvaldehyde-bis(4-methyl-3-thiosemicarbazone), dispersed in a pharmaceutical suppository base.

6. A method for combined oral and vaginal treatment of T richomonas vaginalis and T richomonas foetus infections in human females and cows which comprises: concomitant administration of (1) an oral composition containing, as the pirmary active ingredient, from about to about 225 mg. of a compound of the formula wherein R is a lower alkyl of 1 through 4 carbon atoms.

References Cited by the Examiner FOREIGN PATENTS 1/ 1953 Austria.

OTHER REFERENCES French et a1.: Cancer Research, vol. 18, No. 11, Decernber 1958, pp. 12901300.

JULIAN S. LEVITT, Primary Examiner.

WILLIAM B. KNIGHT, MORRIS O. WOLK, IRVING MARCUS, Examiners.

R. MANNING, R. M. GRANIEWSKI, VERA C.

CLARKE, Assistant Examiners. 

1. A METHOD FOR TREATING TRICHOMONAS VAGINALIS AND TRICHOMONAS FOETUS INFECTIONS IN HUMANS AND CATTLE WHICH COMPRISES: ORALLY ADMINISTERING A COMPOSITION CONTAINING, AS THE PRIMARY ACTIVE INGREDIENT, A COMPOUND OF THE FORMULA 